CAJ | 학술논문

目的探讨罗格列酮(RGZ)与全反式维甲酸(ATRA)在体内外对骨髓瘤细胞生长的影响及其作用途径.方法用RG2和ATRA单独或联合处理多发性骨髓瘤细胞系U266和RPMI 8226细胞,采用3H-TdR掺入法检测细胞增殖变化;Annexin-V和PI双染后采用流式细胞术检测细胞凋亡;用半定量RT-PCR方法检测凋亡抑制基因FLIP、XIAP及survivin mRNA表达变化;采用荧光CaspACE试剂盒检测caspase-3活性变化;采用裸鼠体内成瘤试验观察骨髓瘤细胞在动物体内生长情况.结果 RGZ对骨髓瘤细胞具有增殖抑制作用,且呈剂量依赖关系(U266组r=0.991,P<0.01;RPMI8226组r=0.961,P<0.01);与RGZ单用组比较,ATRA与RGZ联合对骨髓瘤细胞的增殖抑制作用更明显(U266组P<0.001;RPMI8226组P<0.01);10μmol/L的RGZ处理组U266细胞凋亡率为(9.8±1.7)%,RPMI8226细胞凋亡率为(10.7±3.3)%,且诱导凋亡作用呈时间、剂量依赖性,ATRA与RGZ联合应用时,凋亡细胞的比例较相应的RGZ单独处理组显著升高(P值均<0.01);RGZ能抑制FLIP、XIAP及survivin mRNA的表达,ATRA能协同RGZ进一步抑制FLIP、XIAP及survivin mRNA的表达;经RGZ处理的骨髓瘤细胞caspase-3活性显著增加,RGZ与ATRA联合处理后caspase-3活性较RGZ处理组显著增加(P<0.01);RGZ可明显抑制骨髓瘤细胞在裸鼠体内的生长(P<0.001),并且ATRA与RGZ联合能增强RGZ在裸鼠体内对骨髓瘤细胞生长的抑制作用(P<0.01).结论 RGZ能通过抑制FLIP、XIAP及survivin的表达、促进caspase-3的活化从而诱导细胞凋亡,抑制细胞增殖.ATRA能增强RGZ对骨髓瘤细胞的上述作用,两者联合具有协同效应.
목적 탐토라격렬동(RGZ)여전반식유갑산(ATRA)재체내외대골수류세포생장적영향급기작용도경.방법 용RG2화ATRA단독혹연합처리다발성골수류세포계U266화RPMI 8226세포,채용3H-TdR참입법검측세포증식변화;Annexin-V화PI쌍염후채용류식세포술검측세포조망;용반정량RT-PCR방법검측조망억제기인FLIP、XIAP급survivin mRNA표체변화;채용형광CaspACE시제합검측caspase-3활성변화;채용라서체내성류시험관찰골수류세포재동물체내생장정황.결과 RGZ대골수류세포구유증식억제작용,차정제량의뢰관계(U266조r=0.991,P<0.01;RPMI8226조r=0.961,P<0.01);여RGZ단용조비교,ATRA여RGZ연합대골수류세포적증식억제작용경명현(U266조P<0.001;RPMI8226조P<0.01);10μmol/L적RGZ처리조U266세포조망솔위(9.8±1.7)%,RPMI8226세포조망솔위(10.7±3.3)%,차유도조망작용정시간、제량의뢰성,ATRA여RGZ연합응용시,조망세포적비례교상응적RGZ단독처리조현저승고(P치균<0.01);RGZ능억제FLIP、XIAP급survivin mRNA적표체,ATRA능협동RGZ진일보억제FLIP、XIAP급survivin mRNA적표체;경RGZ처리적골수류세포caspase-3활성현저증가,RGZ여ATRA연합처리후caspase-3활성교RGZ처리조현저증가(P<0.01);RGZ가명현억제골수류세포재라서체내적생장(P<0.001),병차ATRA여RGZ연합능증강RGZ재라서체내대골수류세포생장적억제작용(P<0.01).결론 RGZ능통과억제FLIP、XIAP급survivin적표체、촉진caspase-3적활화종이유도세포조망,억제세포증식.ATRA능증강RGZ대골수류세포적상술작용,량자연합구유협동효응.
Objective To investigate the effects of artificial ligand of peroxisome proliferators activated receptors(PPARs),rosiglitazone(RGZ)and all trans-retinoic acid(ATRA)on human myeloma cell line growth in vitro and in vivo.Methods U266 and RPMI 8226 cells were treated with different concentration of RGZ in the presence or absence of ATRA and the results were studied by3H-TdR thymidine incorporation(for cells proliferation),Annexin V-PI staining and caspase-3 activity assay(for cells apoptosis),RTPCR(for FLIP,XIAP and survivin mRNA expression),and tumor formation test in BALB/c nude mice.Resuits Exposure to RGZ induced proliferation inhibition in a dose-dependent manner in both U266(r=0.991,P<0.01)and RPMI 8226 cells(r=0.961,P<0.01).A combination of RGZ with ATRA could enhalice the inhibition effect(P<0.001 in U266,P<0.01 in RPMI8226).10μmol/L ofRGZ induced apoptosis of(9.8±1.7)%in U266 cells and(10.7±3.3)%in RPMI8226 cells,in a time and dose dependent manner and combined with ATRA intensified the apoptosis induction effects(P<0.01 in both cell lines).The FLIP,XIAP and survivin mRNAs were expressed in both cell lines and their levels decreased significandy after cultured with RGZ.The addition of RGZ+ATRA in the culture further decreased the level.Caspase-3 activitv increased substantiaUy with the increase of RGZ concentration and the addition of RGZ+ATRA in the culture medium showed similar synergism effect on easpase-3 activation(P<0.01).The xenograft of U266 cells in BALB/c nude mice were inhibited by RGZ and so did more by the combination of RGZ and ATRA(P<0.01).Conclusion The down-regulation of FLIP,XIAP and Survivin induced by RGZ Can activate caspase-3.whereby induced apoptosis and proliferation inhibition in myeloma cells.ATRA can enhance these effects of RGZ.